BMP-1, MAMMALIAN TOLLOID AND RELATED DEVELOPMENTAL GENES

BMP-1、哺乳动物 Tolloid 和相关发育基因

• 批准号: 2899897
• 负责人: DANIEL S GREENSPAN
• 金额: $ 24.04万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2899897
• 关键词: RNA splicing; RNase protection assay; SDS polyacrylamide gel electrophoresis; central nervous system; developmental genetics; endopeptidases; enzyme activity; extracellular matrix proteins; gene expression; gene interaction; gene targeting; genetic enhancer element; immunocytochemistry; in situ hybridization; laboratory mouse; mammalian embryology; northern blottings; nucleic acid sequence; polymerase chain reaction; procollagen; protein biosynthesis; protein structure function; restriction fragment length polymorphism; site directed mutagenesis; southern blotting; transforming growth factors

Proposed are studies into the structure, expression, and function of a small group of structurally related genes likely to be important in matrix production and in normal, and perhaps abnormal, development of mammalian embryos. Studies include: 1) Further characterization of the gene (PCOLCE), whose protein product, the procollagen C-proteinase enhancer protein, is involved in biosynthesis of type l collagen, the major component of matrix. Studies will include: A) Determining temporal and spatial distribution of expression of PCOLCE during development and comparison to the distribution of expression of other genes with whose products the enhancer protein might interact (eg. the type J collagen gene COL1A1; the BMP-1/mTld gene, which may encode the type I procollagen C-proteinase; and a novel BMP-1/mTld-like gene recently discovered in the P.I's lab); B) Functional study of the mechanism whereby the enhancer protein stimulates the activity of C-proteinase and assaying whether the enhancer may also stimulate activation of TGF-beta-like proteins: C) Studying the possibility that PCOLCE is up-regulated by TGF- beta. Such studies should provide further insight into the role(s) of the enhancer protein and possible interactions with other proteins in developmental and physiological processes. 2) Definitive study of a gene (BMP-1/mTld) encoding alternatively spliced transcripts for a protein involved in organogenesis of bone (BMP-1) and for a mammalian homologue (mTld) of a protein important in dorsal/ventral patterning of Drosophila embryos (tolloid). Studies will: A) Determine the extent of differential expression of the various alternatively spliced RNA forms in embryonic extraembryonic and adult tissues and distribution of mTld in the central nervous system: B) Demonstrate that the protein products of the various alternatively spliced RNA forms exist and localization of these proteins in developing and adult tissues. and determine whether they may be associated with cell membranes: C) Confirm and extend the original observation that BMP-1 is. or is very closely related to the physiological type I procollagen C-proteinase: D) Confirm and extend the original observation that BMP-1 can directly activate TGF- beta-like molecules: E) Determine possible up-regulation of expression of BMP-1/mTld by TGF-beta; F) Investigate possible enzymatic activities of mTld. 3) Characterization of a new gene encoding a protein product with a domain structure identical to but sequence different than. that of mTld. This gene was recently isolated in the P.I.'s lab from a cDNA library constructed from mRNA of +/- knockout mouse embryos which no longer express BMP-1/mTld but which seem to express compensatory enzymatic activity. Characterization will include determining: A) Full-length coding sequences: B) Chromosomal locations in mouse and human genomes: C) Whether alternative splicing occurs: D) Spatial and temporal patterns of expression in adult and developing tissues; E) Possible functions of the protein product(s): F) Whether levels and/or distribution of expression are changed to compensate for loss of BMP-1/mTld in +/- knockout embryos: G) Role(s) of the new gene in development, determined through creation and study of knockout mice homozygous for null alleles of the new gene.

提出的是对A的结构，表达和功能的研究 小组与结构相关的基因可能在基质中很重要 哺乳动物的生产，正常，也许异常的发展 胚胎。研究包括： 1）基因的进一步表征（PColce），其蛋白质产物， Procollagen C蛋白酶增强蛋白参与生物合成 L型胶原蛋白，矩阵的主要组成部分。研究将包括： a）确定pcolce表达的时间和空间分布 在开发和比较与表达的分布中 其他基因与其产物增强蛋白的产物可能相互作用（例如。 J型胶原基因Col1a1; BMP-1/MTLD基因，可能编码 I型Procollagen C蛋白酶；以及一个新颖的BMP-1/MTLD样基因 在P.I的实验室中发现）； b）机制的功能研究 增强蛋白刺激C蛋白酶和测定的活性 增强子是否还可以刺激类似TGF-beta的激活 蛋白质：c）研究PColce被TGF-上调的可能性 beta。这样的研究应进一步了解 增强蛋白和可能与其他蛋白质相互作用 发展和生理过程。 2）编码剪接的基因（BMP-1/MTLD）的确定性研究 参与骨骼的蛋白质（BMP-1）和 对于在背/腹侧重要的蛋白质的哺乳动物同源物（MTLD） 果蝇胚胎（Tolloid）的图案。研究将：a）确定 各种剪接RNA的差分表达的程度 胚胎外囊性和成人组织中的形式以及分布 中枢神经系统中的MTLD：b）证明蛋白质 存在各种剪接的RNA形式的产品，并且 这些蛋白质在发育和成人组织中的定位。和 确定它们是否可能与细胞膜相关：c）确认 并扩展了BMP-1的原始观察结果。或非常紧密 与生理I型Procollagen C蛋白酶有关：D）确认 并扩展了BMP-1可以直接激活TGF-的原始观察结果 β类分子：e）确定可能表达的上调 tgf-beta的BMP-1/MTLD; f）研究可能的酶活性 mtld。 3）表征编码具有域蛋白质产品的新基因 结构与序列相同，但序列不同。 MTLD。这 最近，基因从cDNA文库中分离出来的P.I.实验室 由+/-敲除小鼠胚胎的mRNA构建 表示BMP-1/MTLD，但似乎表达了补偿性酶促的 活动。表征将包括确定：a）全长编码 序列：b）小鼠和人类基因组中的染色体位置：c）是否是否 发生替代剪接：d） 在成人和发展组织中的表达； e） 蛋白质产物：f）表达的水平和/或分布 更改以补偿+/-敲除胚胎中BMP-1/MTLD的损失： g）新基因在开发中的作用，通过创造和 对新基因的无效等位基因纯合子的基因敲除小鼠的研究。

项目成果

Post-translational proteolytic processing of procollagen C-terminal proteinase enhancer releases a metalloproteinase inhibitor.

前胶原 C 末端蛋白酶增强剂的翻译后蛋白水解加工释放金属蛋白酶抑制剂。

• DOI: 10.1074/jbc.275.2.1384
• 发表时间: 2000
• 期刊: The Journal of biological chemistry
• 作者: Mott,JD;Thomas,CL;Rosenbach,MT;Takahara,K;Greenspan,DS;Banda,MJ
• 通讯作者: Banda,MJ

Structural organization and expression patterns of the human and mouse genes for the type I procollagen COOH-terminal proteinase enhancer protein.

人类和小鼠 I 型前胶原 COOH 末端蛋白酶增强蛋白基因的结构组织和表达模式。

• DOI: 10.1006/geno.1998.5663
• 发表时间: 1999
• 期刊: Genomics.
• 作者: Scott,IC;Clark,TG;Takahara,K;Hoffman,GG;Greenspan,DS
• 通讯作者: Greenspan,DS

Coding sequence and expression patterns of mouse chordin and mapping of the cognate mouse chrd and human CHRD genes.

• DOI: 10.1006/geno.1998.5474
• 发表时间: 1998-09
• 期刊: Genomics
• 影响因子: 4.4
• 作者: W. Pappano;Ian C. Scott;Timothy G. Clark;Roger L. Eddy;T. B. Shows;D. Greenspan